Detection of Toxoplasma gondii oocysts in fresh vegetables and berry fruits
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.18/7639 |
Resumo: | Background: Toxoplasma gondii is the third most important contributor to health burden caused by food-borne illness. Ingestion of tissue cysts from undercooked meat is an important source of horizontal transmission to humans. However, there is an increasing awareness of the consumption of fresh fruit and vegetables, as a possible source for oocyst transmission, since this stage of the parasite can persist and remain infective in soil and water for long time. Herein, we outline findings related with detection of T. gondii oocysts in vegetables and berry fruits, which are usually raw consumed. The procedure includes the estimation of the number of oocysts. Methods: Food samples were collected from local producers and supermarket suppliers. Toxoplasma gondii oocysts were concentrated after washing the samples by applying high resolution water filtration and immunomagnetic separation (method 1623.1: EPA 816-R-12-001-Jan 2012), in order to (i) remove potential Cryptosporidium spp. oocysts and Giardia spp. cysts present in the samples; and (ii) select T. gondii oocysts. Toxoplasma gondii oocyst detection and an estimation of their numbers was performed by conventional PCR and real time qPCR, using specific primers for a 183-bp sequence of the T. gondii repetitive DNA region. All PCR-positive DNA samples were purified and sequenced. Restriction enzyme digestion with EcoRV endonuclease confirmed the presence of the T. gondii DNA fragment. In addition, the presence of the parasite was observed by fluorescent microscopy, taking advantage of the oocysts autofluorescence under UV light. Results: Forty percent of the analysed samples (95% CI: 25.5–56.5%) presented the expected PCR and digested DNA fragments. These fragments were confirmed by sequencing. Microscopic autofluorescence supported the presence of T. gondii-like oocysts. The estimated mean (± SE) oocyst concentration was 23.5 ± 12.1 oocysts/g, with a range of 0.6–179.9 oocysts/g. Conclusions: Our findings provide relevant evidence of contamination of fresh vegetables and berry fruits with T. gondii oocysts. |
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Detection of Toxoplasma gondii oocysts in fresh vegetables and berry fruitsFruitOocystsPortugalRaw FoodsSpainToxoplasmaVegetablesFood ParasitologyInfecções GastrointestinaisBackground: Toxoplasma gondii is the third most important contributor to health burden caused by food-borne illness. Ingestion of tissue cysts from undercooked meat is an important source of horizontal transmission to humans. However, there is an increasing awareness of the consumption of fresh fruit and vegetables, as a possible source for oocyst transmission, since this stage of the parasite can persist and remain infective in soil and water for long time. Herein, we outline findings related with detection of T. gondii oocysts in vegetables and berry fruits, which are usually raw consumed. The procedure includes the estimation of the number of oocysts. Methods: Food samples were collected from local producers and supermarket suppliers. Toxoplasma gondii oocysts were concentrated after washing the samples by applying high resolution water filtration and immunomagnetic separation (method 1623.1: EPA 816-R-12-001-Jan 2012), in order to (i) remove potential Cryptosporidium spp. oocysts and Giardia spp. cysts present in the samples; and (ii) select T. gondii oocysts. Toxoplasma gondii oocyst detection and an estimation of their numbers was performed by conventional PCR and real time qPCR, using specific primers for a 183-bp sequence of the T. gondii repetitive DNA region. All PCR-positive DNA samples were purified and sequenced. Restriction enzyme digestion with EcoRV endonuclease confirmed the presence of the T. gondii DNA fragment. In addition, the presence of the parasite was observed by fluorescent microscopy, taking advantage of the oocysts autofluorescence under UV light. Results: Forty percent of the analysed samples (95% CI: 25.5–56.5%) presented the expected PCR and digested DNA fragments. These fragments were confirmed by sequencing. Microscopic autofluorescence supported the presence of T. gondii-like oocysts. The estimated mean (± SE) oocyst concentration was 23.5 ± 12.1 oocysts/g, with a range of 0.6–179.9 oocysts/g. Conclusions: Our findings provide relevant evidence of contamination of fresh vegetables and berry fruits with T. gondii oocysts.This study was supported by the Horizon 2020 project SafeConsume (Grant Agreement No. 727580).BMCRepositório Científico do Instituto Nacional de SaúdeMarques, Cláudia S.Sousa, SusanaCastro, Antónioda Costa, José Manuel Correia2021-04-05T16:18:03Z2020-04-082020-04-08T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.18/7639engParasit Vectors. 2020 Apr 8;13(1):180. doi: 10.1186/s13071-020-04040-2.1756-330510.1186/s13071-020-04040-2info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-20T15:42:07Zoai:repositorio.insa.pt:10400.18/7639Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:42:16.675854Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Detection of Toxoplasma gondii oocysts in fresh vegetables and berry fruits |
title |
Detection of Toxoplasma gondii oocysts in fresh vegetables and berry fruits |
spellingShingle |
Detection of Toxoplasma gondii oocysts in fresh vegetables and berry fruits Marques, Cláudia S. Fruit Oocysts Portugal Raw Foods Spain Toxoplasma Vegetables Food Parasitology Infecções Gastrointestinais |
title_short |
Detection of Toxoplasma gondii oocysts in fresh vegetables and berry fruits |
title_full |
Detection of Toxoplasma gondii oocysts in fresh vegetables and berry fruits |
title_fullStr |
Detection of Toxoplasma gondii oocysts in fresh vegetables and berry fruits |
title_full_unstemmed |
Detection of Toxoplasma gondii oocysts in fresh vegetables and berry fruits |
title_sort |
Detection of Toxoplasma gondii oocysts in fresh vegetables and berry fruits |
author |
Marques, Cláudia S. |
author_facet |
Marques, Cláudia S. Sousa, Susana Castro, António da Costa, José Manuel Correia |
author_role |
author |
author2 |
Sousa, Susana Castro, António da Costa, José Manuel Correia |
author2_role |
author author author |
dc.contributor.none.fl_str_mv |
Repositório Científico do Instituto Nacional de Saúde |
dc.contributor.author.fl_str_mv |
Marques, Cláudia S. Sousa, Susana Castro, António da Costa, José Manuel Correia |
dc.subject.por.fl_str_mv |
Fruit Oocysts Portugal Raw Foods Spain Toxoplasma Vegetables Food Parasitology Infecções Gastrointestinais |
topic |
Fruit Oocysts Portugal Raw Foods Spain Toxoplasma Vegetables Food Parasitology Infecções Gastrointestinais |
description |
Background: Toxoplasma gondii is the third most important contributor to health burden caused by food-borne illness. Ingestion of tissue cysts from undercooked meat is an important source of horizontal transmission to humans. However, there is an increasing awareness of the consumption of fresh fruit and vegetables, as a possible source for oocyst transmission, since this stage of the parasite can persist and remain infective in soil and water for long time. Herein, we outline findings related with detection of T. gondii oocysts in vegetables and berry fruits, which are usually raw consumed. The procedure includes the estimation of the number of oocysts. Methods: Food samples were collected from local producers and supermarket suppliers. Toxoplasma gondii oocysts were concentrated after washing the samples by applying high resolution water filtration and immunomagnetic separation (method 1623.1: EPA 816-R-12-001-Jan 2012), in order to (i) remove potential Cryptosporidium spp. oocysts and Giardia spp. cysts present in the samples; and (ii) select T. gondii oocysts. Toxoplasma gondii oocyst detection and an estimation of their numbers was performed by conventional PCR and real time qPCR, using specific primers for a 183-bp sequence of the T. gondii repetitive DNA region. All PCR-positive DNA samples were purified and sequenced. Restriction enzyme digestion with EcoRV endonuclease confirmed the presence of the T. gondii DNA fragment. In addition, the presence of the parasite was observed by fluorescent microscopy, taking advantage of the oocysts autofluorescence under UV light. Results: Forty percent of the analysed samples (95% CI: 25.5–56.5%) presented the expected PCR and digested DNA fragments. These fragments were confirmed by sequencing. Microscopic autofluorescence supported the presence of T. gondii-like oocysts. The estimated mean (± SE) oocyst concentration was 23.5 ± 12.1 oocysts/g, with a range of 0.6–179.9 oocysts/g. Conclusions: Our findings provide relevant evidence of contamination of fresh vegetables and berry fruits with T. gondii oocysts. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-04-08 2020-04-08T00:00:00Z 2021-04-05T16:18:03Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.18/7639 |
url |
http://hdl.handle.net/10400.18/7639 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Parasit Vectors. 2020 Apr 8;13(1):180. doi: 10.1186/s13071-020-04040-2. 1756-3305 10.1186/s13071-020-04040-2 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
BMC |
publisher.none.fl_str_mv |
BMC |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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